title: The DNT-EST: a predictive embryonic stem cell-based assay for developmental neurotoxicity testing in vitro.
abstract: As the developing brain is exquisitely vulnerable to chemical disturbances, testing for developmental neurotoxicity of a substance is an important aspect of characterizing its tissue specific toxicity. Mouse embryonic stem cells (mESCs) can be differentiated toward a neural phenotype, and this can be used as a model for early brain development. We developed a new in vitro assay using mESCs to predict adverse effects of chemicals and other compounds on neural development - the so-called DNT-EST. After treatment of differentiating stem cells for 48h or 72h, at two key developmental stages endpoint for neural differentiation, viability, and proliferation were assessed. As a reference, we similarly treated undifferentiated stem cells 2 days after plating for 48h or 72h in parallel to the differentiating stem cells. Here, we show that chemical testing of a training set comprising nine substances (six substances of known developmental toxicity and three without specific developmental neurotoxicity) enabled a mathematical prediction model to be formulated that provided 100% predictivity and accuracy for the given substances, including in leave-one-out cross-validation. The described test method can be performed within two weeks, including data analysis, and provides a prediction of the developmental neurotoxicity potency of a substance.
authors.affiliation: German Federal Institute for Risk Assessment (BfR), Department of Experimental Toxicology and ZEBET, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Hayess
authors.forename: Christian
authors.initials: C
authors.lastname: Riebeling
authors.forename: Ralph
authors.initials: R
authors.lastname: Pirow
authors.forename: Matthias
authors.initials: M
authors.lastname: Steinfath
authors.forename: Dana
authors.initials: D
authors.lastname: Sittner
authors.forename: Birgitta
authors.initials: B
authors.lastname: Slawik
authors.forename: Andreas
authors.initials: A
authors.lastname: Luch
authors.forename: Andrea E M
authors.initials: AE
authors.lastname: Seiler
mesh_terms: D000465:Algorithms; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003627:Data Interpretation, Statistical; D016002:Discriminant Analysis; D053595:Embryonic Stem Cells / Q000187:drug effects*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D058953:Neural Stem Cells / Q000187:drug effects; D009474:Neurons / Q000187:drug effects; D020258:Neurotoxicity Syndromes / Q000473:pathology*; D009498:Neurotoxins / Q000633:toxicity*; D016716:PC12 Cells; D011237:Predictive Value of Tests; D051381:Rats; D018675:Toxicity Tests / Q000379:methods*; D014404:Tubulin / Q000378:metabolism
chemical_list: D009498:Neurotoxins; D014404:Tubulin; D019950:Mitogen-Activated Protein Kinase 1
keywords: Alternative method to animal experiments; Developmental neurotoxicity; Embryonic stem cells; In vitro developmental neurotoxicity testing